We compared simultaneous dual-radionuclide (DR) stress and rest myocardial perfusion imaging (MPI) with a novel solid-state cardiac camera and a conventional SPECT camera with separate stress and rest acquisitions. Of 27 consecutive patients recruited, 24 (64.5±11.8 years of age, 16 men) were injected with 74 MBq of 201Tl (rest) and 250 MBq 99mTc-MIBI (stress). Conventional MPI acquisition times for stress and rest are 21 min and 16 min, respectively. Rest 201Tl for 6 min and simultaneous DR 15-min list mode gated scans were performed on a D-SPECT cardiac scanner. In 11 patients DR D-SPECT was performed first and in 13 patients conventional stress 99mTc-MIBI SPECT imaging was performed followed by DR D-SPECT. The DR D-SPECT data were processed using a spill-over and scatter correction method. DR D-SPECT images were compared with rest 201Tl D-SPECT and with conventional SPECT images by visual analysis employing the 17-segment model and a five-point scale (0 normal, 4 absent) to calculate the summed stress and rest scores. Image quality was assessed on a four-point scale (1 poor, 4 very good) and gut activity was assessed on a four-point scale (0 none, 3 high). Conventional MPI studies were abnormal at stress in 17 patients and at rest in 9 patients. In the 17 abnormal stress studies DR D-SPECT MPI showed 113 abnormal segments and conventional MPI showed 93 abnormal segments. In the nine abnormal rest studies DR D-SPECT showed 45 abnormal segments and conventional MPI showed 48 abnormal segments. The summed stress and rest scores on conventional SPECT and DR D-SPECT were highly correlated ( r=0.9790 and 0.9694, respectively). The summed scores of rest 201Tl D-SPECT and DR-DSPECT were also highly correlated ( r=0.9968, p<0.0001 for all). In six patients stress perfusion defects were significantly larger on stress DR D-SPECT images, and five of these patients were imaged earlier by D-SPECT than by conventional SPECT. Fast and high-quality simultaneous DR MPI is feasible with D-SPECT in a single imaging session with comparable diagnostic performance and image quality to conventional SPECT and to a separate rest 201Tl D-SPECT acquisition.